天等1号辣椒遗传转化再生体系的研究

对天等1号辣椒的子叶、带柄子叶、带柄真叶和F lam ingo-b ill外植体进行了遗传转化,比较不同激素及其水平对不同外植体芽分化的影响,以及外植体对卡那霉素的敏感性。结果表明,仅有以F lam ingo-b ill为外植体所形成的愈伤组织,能在培养基M S ZT 1.0m g/L IAA 0.2m g/L A gNO32.0m g/L上诱导芽的分化(63%的分化率),在培养基M S ZT 1.0m g/L IAA 0.2m g/L A gNO32.0m g/L GA32.0m g/L上使诱导芽伸长(15%的芽伸长率)。抗性筛选最适卡那霉素浓度为50m g/L。通过根癌农杆菌介导法转化,共获得33株抗性再生苗。PCR和PCR-Sou thern杂交检测结果表明,94%的抗性植株检测到报告基因nptII片段。

Study on genetic regeneration and transformation of pepper variety Tiandeng No.1

Four explants including cotyledon,cotyledon with petiole, euphylla with petiole and Flamingo-bill of the pepper cultivar 'Tiandeng No.1' were used for genetic transformation to compare the effects of hormone types, hormone combinations and levels on the buds induction of different explants,and know about the sensitivity of different explants to kanamycin.The results showed that only the callus induced from Flamingo-bill explants could induce the buds differentiation on the medium MS ZT 1.0 mg/L IAA 0.2 mg/L AgNO_3 2.0 mg/L, and the inducing rate was 63%.It also could induce the elongation of buds on medium MS ZT 1.0 mg/L IAA 0.2 mg/L AgNO_3 2.0 mg/L GA_3 2.0 mg/L,and the elongation rate was 15%.The optimal concentration of antibiotic kanamycin for screening the transformed Flamingo-bill explants was 50mg/L.By Agrobacterium tumefaciens-mediated transformation,33 marker-positive regenerated pepper plantlets were obtained,and 94% of them were confirmed to carry the reporter gene-nptII by PCR and PCR-Southern analysis.